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Retention behaviour and fluorimetric detection of procaine hydrochloride using carboxymethyl-beta-cyclodextrin as an additive in reversed-phase liquid chromatography.

The retention behaviour of procaine hydrochloride on an Alltima octadecyl silica (C18) column, with a mobile phase containing negatively charged carboxymethyl-beta-cyclodextrin (CM-beta-CD), influenced by a combination of hydrophobic and electrostatic interactions was systematically investigated. Various factors affecting the retention of procaine on the C18 column such as the concentration of CM-beta-CD, pH and the methanol percentage in the mobile phase, were studied. An equation was applied to estimate the apparent binding constant of the CM-beta-CD-procaine inclusion complex as an aid for understanding the retention mechanism. The first analytical application of CM-beta-CD as a mobile phase additive for the determination of procaine was developed. The calibration curve was linear in the range 22-1360 ng ml(-1) with an RSD of 2.1%. The detection limit based on 3sigma was 1 ng ml(-1) with fluorimetric detection at the excitation and emission wavelengths of 305 nm and 350 nm, respectively. The limit of quantitation based on 10sigma was 22 ng ml(-1). The proposed method has been successfully applied to real sample analysis.

Chromatography, Liquid↗

A method for routine analysis of recombinant immunoglobulins (rIgGs) by capillary isoelectric focusing (cIEF).

A capillary isoelectric focusing (cIEF) method was developed for routine analysis of recombinant immunoglobulins (rlgGs). The cIEF method used a dimethyl siloxane-coated capillary and a separation matrix of 2% ampholytes in 0.4% methylcellulose (MC). The rIgGs, and internal pI marker protein standards, were mixed with carrier ampholyte in MC, focused using high voltage, and then the protein bands were mobilized past a UV detector by simultaneous application of low pressure and voltage. Qualitatively and quantitatively equivalent rIgG focusing profiles were obtained via cIEF and gel-based IEF, with individual isoform peak area percentages and calculated peak pI values being comparable for the same samples. Linear relationships were obtained for peak area response versus sample concentration, and for the pI gradient developed between the internal pI marker standards. The relative standard deviation (RSD) in rIgG peak areas was less than 2% intra-day and less than 8% inter-day (72 h). The RSD for the mobilization times of rIgG peaks was less than 1% intra-day and less than 3% inter-day (72 h). There was no observed decrease in the performance of the capillary over 150 analyses. cIEF offers several important advantages over gel IEF, e.g. direct, quantitative detection of proteins by intrinsic UV absorbance at 280 nm, rapid analyses ( < or = 30 min), capability of automation, and one-step, electronic data analysis and archival. These data demonstrate the superiority of the cIEF method for routine analysis of rIgGs.

Buffers↗

Impact of finer grid resolution on the spatial distribution of vehicle emissions inventories.

The ability to model air quality dispersion at increasingly smaller resolutions requires a concomitant improvement in the resolution of the gridded mobile source emissions used as input to these models. Two methods are currently used to allocate mobile emissions to grids; because of the limitations associated with these methods, their application is usually restricted to coarser grid resolutions. This paper uses a new mobile emissions inventory model (UCDrive) to explore the spatial distribution of mobile source emissions using finer grid cell resolutions. Our results indicate that the new model improves the precision of the spatial allocation of mobile source emissions, which in turn improves our ability to identify and implement pollutant control strategies.

Air Pollutants↗

Using software agents to maintain autonomous patient registries for clinical research.

A software agent is an application that can function in an autonomous and intelligent fashion. We have used mobile software agents to maintain clinicians' patient research databases (patient registries). Agents were used to acquire data from the clinician and place it into the registries, copy data from hospital databases into the registries, and report data from the registries. The agents were programmed with the intelligence to navigate through complex network security, interact with legacy systems, and protect themselves from various forms of failure at multiple levels. To maximize the separation between our system and the hospital information infrastructure we often used Java, a platform-independent language, to program and distribute our software agents. By using mobile agents, we were able to distribute the computing time required by these applications to underutilized host machines upon which the registries could be maintained.

Databases as Topic↗

Rollover-free navigation for a mobile agent in an unstructured environment.

This paper introduces a navigation method for a teleoperated mobile agent (or robot) moving in an unstructured environment that includes unknown obstacles and uneven terrain, based on a guided-navigation algorithm (GNA) and a rollover-prevention algorithm (RPA). Although the mobile agent is primarily driven by an operator at a remote site, it reacts autonomously for avoiding collision with obstacles and for preventing rollover when it suspects/detects possible collision or rollover. The autonomous reactive motion is normally unexpected, thus there exists the inconsistency between the intended motion and the controlled motion of the agent from the operator. A force-reflection technique utilizing a force-feedback joystick is developed to manipulate this inconsistency. To verify the feasibility and effectiveness of the proposed navigation method, experiments with the Robot for Hazardous Application-Double Tracks (ROBHAZ-DT) (actual mobile agent) are successfully carried out.

Artificial Intelligence↗

Trypanosoma brucei: trypanosome strain typing using PCR analysis of mobile genetic elements (MGE-PCR).

We describe the development of a single-primer amplification system, which uses the trypanosomal mobile genetic element RIME as a molecular marker for the differentiation of Trypanosoma brucei stocks. Using a well-characterised set of T. brucei stocks from southeast Uganda, Kenya and Zambia, we have evaluated the application of this technique, termed MGE-PCR (mobile genetic element PCR) for the typing of trypanosome strains. The technique revealed considerable variation between stocks and was sufficiently specific to amplify trypanosomal DNA in the presence of host DNA. The results showed a clear distinction between human-infective and non-human-infective stocks. Comparative studies on these stocks using markers for the human serum resistance associated (SRA) gene, which identifies human-infective stocks, demonstrated complete agreement between MGE-PCR derived groups and human-infectivity status. Furthermore, MGE-PCR detects high levels of variability within the T. b. brucei and T. b. rhodesiense groups and is therefore a powerful discriminatory tool for tracking individual T. brucei genotypes and strains.

Animals↗

Potential role for a guanine nucleotide regulatory protein in chemoattractant receptor mediated polyphosphoinositide metabolism, Ca++ mobilization and cellular responses by leukocytes.

Islet activating protein from Bordetella pertussis toxin which ribosylates certain guanine nucleotide regulatory proteins causes a marked reduction of chemoattractant-elicited responses such as chemotaxis, O2 production and cAMP elevations in human polymorphonuclear leukocytes. The toxin appears to exert its effects by preventing the rapid breakdown of phosphatidylinositol 4,5-bisphosphate induced by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine, thereby inhibiting the increase in intracellular [Ca++] which normally follows chemoattractant stimulation. Responses of leukocytes exposed to Concanavalin A, the Ca++ ionophore A23187, or phorbol myristate acetate were not affected by the toxin. Thus the chemoattractant receptor appears to be coupled to a phosphoinositide specific phospholipase C through a guanine nucleotide regulatory protein. We propose that this complex of receptor-guanine nucleotide regulatory protein-phospholipase C may be applicable to the class of receptors which mobilize intracellular Ca++ by stimulating polyphosphoinositide breakdown.

Bacterial Toxins↗

REDOR-based heteronuclear dipolar correlation experiments in multi-spin systems: rotor-encoding, directing, and multiple distance and angle determination.

We review a variety of recently developed 1H-X heteronuclear recoupling techniques, which rely only on the homonuclear decoupling efficiency of very-fast magic-angle spinning. All these techniques, which are based on the simple rotational-echo, double-resonance (REDOR) approach for heteronuclear recoupling, are presented in a common context. Advantages and possibilities with respect to the complementary application of conventionally X and 1H-inversely detected variants are discussed in relation to the separability and analysis of multiple couplings. We present an improved and more sensitive approach to the determination of 1H-X dipolar couplings by spinning-sideband analysis, termed REREDOR, which is applicable to XHn groups in rigid and mobile systems and bears some similarity to more elaborate separated local-field methods. The estimation of medium-range 1H-X distances by analyzing signal intensities in two-dimensional REDOR correlation spectra in a model-free way is also discussed. More specifically, we demonstrate the possibility of combined distance and angle determination in H-X-H or X-H-X three-spin systems by asymmetric recoupling schemes and spinning-sideband analysis. Finally, an 1H-X correlation experiment is introduced which accomplishes high sensitivity by inverse (1H) detection and is therefore applicable to samples with 15N in natural abundance.

Alanine↗

Community ambulation after stroke: how important and obtainable is it and what measures appear predictive?

OBJECTIVES: To assess how important community ambulation is to stroke survivors and to assess the relation between the level of community ambulation achieved and other aspects of mobility. DESIGN: A multicenter observational survey. SETTING: Community setting in New Zealand. PARTICIPANTS: One hundred fifteen stroke survivors living at home were referred from physical therapy (PT) services at 3 regional hospitals at the time of discharge and were assessed within 1 week after returning home. Another 15 people with stroke who did not require further PT when discharged were assessed within 2 weeks after they returned home to provide insight into community ambulation status for those without mobility impairment, as recognized by health professionals. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Self-reported levels of community ambulation ascertained by questionnaire, gait velocity (m/min), Functional Ambulation Categories (FAC) score, and Rivermead Mobility Index (RMI) score. RESULTS: Mean gait velocity for the participants was 53.9 m/min (95% confidence interval [CI], 52.3-61.1); mean treadmill distance was 165.5 m (95% CI, 141.6-189.5); median RMI score was 14; and median FAC score was 6. Mobility scores for the 15 people who did not require PT were within the normal range. Based on self-reported levels of ambulation, 19 (14.6%) participants were unable to leave the home unsupervised, 22 (16.9%) were walking as far as the letterbox, 10 (7.6%) were limited to walking within their immediate environment, and 79 (60.7%) could access shopping malls and/or places of interest. Participants with different levels of community ambulation showed a significant difference in gait velocity (P<.001). The ability to "get out and about" in the community was considered to be either essential or very important by 97 subjects (74.6%). CONCLUSIONS: Community ambulation is a meaningful outcome after stroke. However, despite good mobility outcomes on standardized measures for this cohort of home-dwelling stroke survivors, nearly one third were not getting out unsupervised in the community. Furthermore, gait velocity may be a measure that discriminates between different categories of community ambulation. These findings may have implications for PT practice for people with mobility problems after stroke.

Aged↗

Transfer of phloem-mobile substances from the host plants to the holoparasite Cuscuta sp.

During the development of the haustorium, searching hyphae of the parasite and the host parenchyma cells are connected by plasmodesmata. Using transgenic tobacco plants expressing a GFP-labelled movement protein of the tobacco mosaic virus, it was demonstrated that the interspecific plasmodesmata are open. The transfer of substances in the phloem from host to the parasite is not selective. After simultaneous application of (3)H-sucrose and (14)C-labelled phloem-mobile amino acids, phytohormones, and xenobiotica to the host, corresponding percentages of the translocated compounds are found in the parasite. An open continuity between the host phloem and the Cuscuta phloem via the haustorium was demonstrated in CLSM pictures after application of the phloem-mobile fluorescent probes, carboxyfluorescein (CF) and hydroxypyrene trisulphonic acid (HPTS), to the host. Using a Cuscuta bridge (14)C-sucrose and the virus PVY(N) were transferred from one host plant to the another. The results of translocation experiments with labelled compounds, phloem-mobile dyes and the virus should be considered as unequivocal evidence for a symplastic transfer of phloem solutes between Cuscuta species and their compatible hosts.

Amino Acids↗

Does 12 weeks of regular standing prevent loss of ankle mobility and bone mineral density in people with recent spinal cord injuries?

The purpose of this study was to determine the effects of a 12-week standing program on ankle mobility and femur bone mineral density in patients with lower limb paralysis following recent spinal cord injury. An assessor-blinded within-subject randomised controlled trial was undertaken. Twenty patients with lower limb paralysis following a recent spinal cord injury were recruited. Subjects stood weight-bearing through one leg on a tilt-table for 30 minutes, three times each week for 12 weeks. By standing on one leg a large dorsiflexion stretch was applied to the ankle and an axial load was applied to the bones of the weight-bearing leg. Ankle mobility and femur bone mineral density of both legs were measured at the beginning and end of the study. Ankle mobility (range of motion) was measured with the application of a 17 Nm dorsiflexion torque. Femur bone mineral density was measured using dual energy X-ray absorptiometry (DEXA). The effect of standing was estimated from the difference between legs in mean change of ankle mobility and femur bone mineral density. The results indicated a mean treatment effect on ankle mobility of 4 degrees (95% CI 2 to 6 degrees) and on femur bone mineral density of 0.005 g/cm(2) (95% CI -0.015 to 0.025 g/cm(2)). Tilt-table standing for 30 minutes, three times per week for 12 weeks has a small effect on ankle mobility, and little or no effect on femur bone mineral density. It is unclear whether clinicians and patients would consider such effects to be clinically worthwhile.

Adult↗

Staging functional independence validity and applications.

OBJECTIVES: To establish construct and predictive validity of the activities of daily living, sphincter-management, mobility, and executive-function (ASME) staging system of functional independence and to describe potential applications. DESIGN: National data were used to validate the stages. Sensitivity to functional change was addressed by comparing patients' stages at discharge to their stages at admission. Construct validity was assessed by examining stage distribution differences among groups of patients classified by impairment category. Predictive validity was determined by the degree to which high stages at admission predict discharge to the community and low stages at admission predict serious morbidity or mortality. SETTING: Inpatient rehabilitation facilities. PARTICIPANTS: Data from 231,686 individuals discharged from 560 US inpatient rehabilitation facilities in 1995. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Stage at rehabilitation discharge, community discharge, acute hospital discharge, or death while in rehabilitation. RESULTS: More than 90% of patients were discharged at a higher stage in at least 1 domain while not declining in any other domain. The odds (95% confidence interval) of community discharge for patients admitted at the highest compared with the lowest stage varied from 2.1 (1.9-2.2) to 3.1 (2.9-3.2), depending on the domain. The odds of acute hospitalization or death were reduced at the highest stages, compared with the lowest, by a factor ranging from .33 (.22-.49) to .65 (.61-.69). CONCLUSIONS: The ASME stages appear valid and sensitive to change. Functional Independence Staging (FIS) has relevance in establishing prognoses and in selecting among therapeutic modalities. FIS will facilitate the identification of meaningful functional goals and interpretable clinical trial endpoints. One of its most important applications could be in the establishment of outcome indicators for quality monitoring under the prospective payment system.

Activities of Daily Living↗

NMR imaging of high-amylose starch tablets. 2. Effect of tablet size.

Carbohydrate polymers are widely used for pharmaceutical applications such as the controlled release of drugs. The swelling and water mobility in high-amylose starch tablets are important parameters to be determined for these applications. They have been studied at different time intervals by nuclear magnetic resonance imaging (NMRI) after the immersion of the samples in water. These tablets have a hydrophilic matrix, which swells anisotropically and forms a hydrogel in water. NMRI shows clearly the anisotropy of the water penetration and the swelling along the radial and axial dimensions of the tablets. Empirical relationships are established to describe the kinetics of water penetration and swelling of the tablets. Results show that water uptake and tablet swelling strongly depend on the size of the tablets. Gravimetric measurements of water uptake were also performed in comparison with the NMRI results.

Absorption↗

Ionic conduction, rectification, and selectivity in single conical nanopores.

Modern track-etching methods allow the preparation of membranes containing a single charged conical nanopore that shows high ionic permselectivity due to the electrical interactions of the surface pore charges with the mobile ions in the aqueous solution. The nanopore has potential applications in electrically assisted single-particle detection, analysis, and separation of biomolecules. We present a detailed theoretical and experimental account of the effects of pore radii and electrolyte concentration on the current-voltage and current-concentration curves. The physical model used is based on the Nernst-Planck and Poisson equations. Since the validity of continuum models for the description of ion transport under different voltages and concentrations is recognized as one of the main issues in the modeling of future applications, special attention is paid to the fundamental understanding of the electrical interactions between the nanopore fixed charges and the mobile charges confined in the reduced volume of the inside solution.

Biological Transport↗

Biomechanic modeling of sit-to-stand to upright posture for mobility assessment of persons with chronic stroke.

OBJECTIVE: To test the suitability of using biomechanic measures associated with a minimum measured input model (MMIM) approach to assess mobility of people with chronic stroke during the execution of a sit-to-stand (STS) to upright posture motor task. DESIGN: Single group, observational. SETTING: Institutional settings in the United States and Italy. PARTICIPANTS: Twenty-nine subjects with chronic unilateral lower-limb impairments and resultant mobility limitations secondary to stroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Manual measurement of lower-limb strength; performance-based tests including repeated chair standing, walking speed, and standing balance; and ground reactions measured with a force platform during STS and upright posture. The ground reactions were fed to a telescopic inverted pendulum model of the musculoskeletal system. Parameters representing the model outputs were compared with performance-based and strength measures to assess, respectively, motor ability and impairment-related changes in subjects' motor strategies. RESULTS: The parameters derived from the model effectively differentiated between motor strategies associated with different performance-based scores, and allowed the identification of relevant difficulties encountered in STS execution. These difficulties could be associated with different strength scores. This was also true for subjects scoring the maximum in both performance-based and strength tests. CONCLUSIONS: The MMIM is a relatively inexpensive and noninvasive approach that enhances mobility assessment of hemiparetic subjects with different motor ability levels. It provides information that correlates well with performance-based and strength scores and, in addition, it allows for subject-specific motor strategy identification.

Aged↗

Comparison of silica-based cyanopropyl and octyl reversed-phase packings for the separation of peptides and proteins.

The performance of a silica-based C8 packing was compared with that of a less hydrophobic, silica-based cyanopropyl (CN) packing during their application to reversed-phase high-performance liquid chromatography (linear trifluoroacetic acid-water to trifluoroacetic acid-acetonitrile gradients) of peptides and proteins. It was found that: (1) the CN column showed excellent selectivity for peptides which varied widely in hydrophobicity and peptide chain length; (2) peptides which could not be resolved easily on the C8 column were widely separated on the CN column; (3) certain mixtures of peptides and small organic molecules which could not be resolved on the C8 column were completely separated on the CN column; (4) impurities arising from solid-phase peptide synthesis were resolved by a wide margin on the CN column, unlike on the C8 column, where these compounds were eluted very close to the peptide product of interest: and (5) specific protein mixtures exhibited superior resolution and peak shape on the CN column compared with the C8 column. The results clearly demonstrate the effectiveness of employing stationary phases of different selectivities (as opposed to the more common optimization protocol of manipulating the mobile phase) for specific peptide and protein applications, an approach underestimated in the past.

Amino Acid Sequence↗

Discontinuous electrophoresis revisited: a review of the process.

The use and development of discontinuous buffer systems for the separation of proteins and nucleic acids are reviewed, and the advantages over a continuous buffer system are presented. Emphasis is given to the more recent applications in DNA separations where discontinuous systems have aided in the resolution and mobility tailoring of DNA fragments. Various DNA size ranges can be separated in short time periods by simple choice of buffer components rather than gel alterations. Guidelines for choosing appropriate buffer systems are offered as well as the application of borate complexes to modify the mobility of DNA. This article reviews the historical development of discontinuous buffers in the electrophoresis of proteins and nucleic acids with emphasis on the characteristics and applicability to nucleic acid separations.

Boric Acids↗

Fate and effects of acrolein.

Acrolein is a highly toxic, reactive, and irritating aldehyde that occurs as a product of organic pyrolysis, as a metabolite of a number of compounds, and as a residue in water when used for the control of aquatic organisms. It is an intermediate in the production of acrylic acid, DL-methionine, and numerous other agents. Its major direct use is as a biocide for the control of aquatic flora and fauna. It is introduced to the environment from a variety of sources, including organic combustion such as automobile exhaust, cigarette smoke, and manufacturing and cooking emissions, as well as direct biocidal applications. Organic combustion from both fixed and mobile sources is the significant source of acrolein in the atmosphere; it represents up to 8% of the total aldehydes generated from vehicles and residential fireplaces and 13% of total atmospheric aldehydes. This reactive aldehyde also occurs in organisms as a metabolite of allyl alcohol, allylamine, spermine, spermidine, and the anticancer drug cyclophosphamide, and as a product of UV radiation of the skin lipid triolein. Furthermore, small amounts are found in foods; when animal or vegetable fats are overheated, however, large amounts are produced. Most human contact occurs during exposure to smoke from cigarettes, automobiles, industrial processes, and structural and vegetation fires. Besides cigarette smoke, occupational exposures are a common mode of human contact, particularly in industries that involve combustion of organic compounds. Firefighters, in particular, are exposed to extremely high levels during the extinguishment and overhaul phases of their work. Water may contain significant levels of the herbicide. It has been found in paper mill and municipal effluents at 20-200 micrograms/L, and at 30 micrograms/L as far as 64 km downstream from the point of application. The USEPA-recommended water quality criteria for freshwater are only 1.2 micrograms/L (24-hr avg) and 2.7 micrograms/L (maximum ceiling). Acrolein is highly reactive, and intercompartmental transport is limited. However, it is eliminated from aqueous environments by volatilization and hydration to beta-hydroxypropanal, after which biotransformation occurs, with a half-life of 7-10 d. The Koc for acrolein is 24, and it is not likely to be retained in soil; activated carbon adsorbs only 30% from solution. Thus, the aldehyde is either leached extensively in moist soil or volatilizes quickly from dry soil. It is eliminated from air by reaction with .OH (half-life, 0.5-1.2 d), NOx (half-life, 16 d), and O3 (half-life, 59 d), as well as by photolysis and wet deposition. As expected from its high water solubility, bioaccumulation is low. Acrolein is highly toxic by all routes of exposure. The respiratory system is the most common target: exposure causes localized irritation, respiratory distress, pulmonary edema, cellular necrosis, and increased susceptibility to microbial diseases. Additionally, acute inhalation studies verify that it is a severe respiratory irritant that affects respiratory rates. Respiratory rate depression may have a protective effect by minimizing vapor inhalation, thereby explaining the subadditive effect of acrolein when combined with the other toxic combustion by-products CO and HCHO. Liquid contact with the skin and eyes causes severe irritation, opaque or cloudy corneas, and localized epidermal necrosis, but no allergic contact dermatitis. The cardiovascular system is affected, resulting in increased blood pressure, platelet aggregation, and quick cessation of beating in perfused rat hearts. It may also inhibit mitochondrial oxidative phosphorylation in the myocardium. Acute LD50s and LC50s are low. Levels are 7-46 mg/kg and 18-750 mg/m3, respectively, in rats; aquatic organisms are affected above 11.4 micrograms/L.(ABSTRACT TRUNCATED)

Acrolein↗